Related Experiment Video
Updated: Apr 5, 2026

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α
Published on: November 2, 2018
Expression, fermentation and purification of a predicted intrinsically disordered region of the transcription factor,
Jenna F DuMond1, Yi He2, Maurice B Burg1
1National Institutes of Health, National Heart, Lung and Blood Institute, Systems Biology Center, Bethesda, MD 20892, United States.
Abstract:
Hypertonicity stimulates Nuclear Factor of Activated T-cells 5 (NFAT5) nuclear localization and transactivating activity. Many transcription factors are known to contain intrinsically disordered regions (IDRs) which become more structured with local environmental changes such as osmolality, temperature and tonicity. The transactivating domain of NFAT5 is predicted to be intrinsically disordered under normal tonicity, and under high NaCl, the activity of this domain is increased. To study the binding of co-regulatory proteins at IDRs a cDNA construct expressing the NFAT5 TAD was created and transformed into Escherichia coli cells. Transformed E. coli cells were mass produced by fermentation and extracted by cell lysis to release the NFAT5 TAD. The NFAT5 TAD was subsequently purified using a His-tag column, cation exchange chromatography as well as hydrophobic interaction chromatography and then characterized by mass spectrometry (MS).
More Related Videos
Related Concept Videos
NF-κB-dependent Signaling Pathway
NF-κB-dependent Signaling Mechanism
The...
Co-activators and Co-repressors
Regulation of Nuclear Protein Sorting
NF-kB-dependent Signaling Pathway
Bacterial Protein Maturation
Regulation of the Unfolded Protein Response

